Biologically relevant orthogonal cytokine/receptor pairs

Abstract

Engineered orthogonal cytokine receptor/ligand pairs, and methods of use thereof, are provided.

What is claimed is: 
     
         1 . A system for selective activation of a receptor in a cell, the system comprising:
 (a) an orthogonal receptor, which does not bind to its native ligand; and   (b) an orthogonal cytokine, which (i) does not bind to its native receptor and (ii) binds to and activates the orthogonal receptor.   
     
     
         2 . The system of  claim 1 , wherein the orthogonal receptor is expressed by a mammalian cell. 
     
     
         3 . The system of  claim 2 , wherein the cell is an immune cell or a stem cell. 
     
     
         4 . The system of  claim 3 , wherein the immune cell is a T cell. 
     
     
         5 . The system of  claim 2 , wherein the cell is a human or mouse cell. 
     
     
         6 . The system of  claim 1 , wherein the orthogonal receptor and orthogonal cytokine are derived from human or mouse proteins. 
     
     
         7 . The system of  claim 1 , wherein the orthogonal receptor is an IL-2 receptor. 
     
     
         8 . The system of  claim 7 , wherein the IL-2 receptor is IL-2N3. 
     
     
         9 . The system of  claim 7 , wherein the orthologous cytokine is IL-2. 
     
     
         10 . The system of  claim 7  wherein the cytokine is IL-15. 
     
     
         11 . The system of  claim 7 , wherein the IL-2 receptor is human CD122 modified at one or more residues selected from R41, R42, Q70, K71, T73, T74, V75, S132, H133, Y134, F135, E136, Q214. 
     
     
         12 . The system of  claim 7 , wherein the IL-2 receptor is mouse CD122 modified at one or more residues selected from R42, F67, Q71, S72, T74, S75, V76, S133, H134, Y135, I136, E137, R215. 
     
     
         13 . The system of  claim 9 , wherein the orthologous IL-2 is human IL-2 modified at one or more residues selected from Q13, L14, E15, H16, L19, D20, Q22, M23, G27, and N88. 
     
     
         14 . The system of  claim 13 , wherein the human IL-2 is modified at one or more residues selected from E15, H16, L19, D20, Q22, and M23. 
     
     
         15 . The system of  claim 9 , wherein the orthologous IL-2 is mouse IL-2, modified at one or residues selected from H27, L28, E29, Q30, M33, D34, Q36, E37, R41, and N103. 
     
     
         16 . The system of  claim 15 , wherein the mouse IL-2 is modified at one or more residues selected from E29, Q30, M33, D34, Q36, and E37. 
     
     
         17 . A composition comprising an orthologous cytokine receptor according to any one of  claim 1 . 
     
     
         18 . A nucleic acid encoding the orthologous receptor of  claim 17 . 
     
     
         19 . An expression vector comprising the nucleic acid of  claim 18 . 
     
     
         20 . A cell genetically engineered to comprise the vector of  claim 19 . 
     
     
         21 . A composition comprising an orthologous cytokine according to  claim 1 . 
     
     
         22 . A nucleic acid encoding the orthologous cytokine of  claim 21 . 
     
     
         23 . An expression vector comprising the nucleic acid of  claim 22 . 
     
     
         24 . A cell genetically engineered to comprise the vector of  claim 23 . 
     
     
         25 . A method of treating an individual, the method comprising introducing a cell according to  claim 20  into the individual, and selectively activating the cell according to the orthologous receptor by contacting with the orthologous cytokine according to  claim 21 . 
     
     
         26 . The method of  claim 25 , wherein the cell is a T cell. 
     
     
         27 . The method of  claim 26 , wherein the individual is treated for cancer. 
     
     
         28 . The method of  claim 26 , wherein the individual is treated for autoimmune disease. 
     
     
         29 . The method of  claim 26 , wherein the individual is treated for infection. 
     
     
         30 . A kit comprising the system of  claim 1 . 
     
     
         31 . The kit according to  claim 30 , comprising a vector or nucleic acid and a cytokine. 
     
     
         32 . A method of obtaining a system according to  claim 1 , comprising the steps of:
 engineering amino acid changes into a native receptor to disrupt binding to the native cytokine;   engineering amino acid changes into the native cytokine at contact residues for receptor binding,   selecting for cytokine orthologs that bind to the ortholog receptor;   discarding ortholog cytokines that bind to the native receptor.   
     
     
         33 . A method of obtaining a system according to  claim 1 , comprising the steps of:
 engineering amino acid changes into a native receptor to disrupt binding to the native cytokine;   engineering amino acid changes into the native cytokine at contact residues for receptor binding,   selecting for receptor orthologs that bind the ortholog cytokine;   discarding ortholog receptors that bind to the native cytokine.